Follicular Atresia, Cell Proliferation, and Anti-Mullerian Hormone in Two Neotropical Primates (Aotus nancymae and Sapajus macrocephalus).

This study evaluated the follicular atresia, cell proliferation, and anti-Mullerian hormone action in Aotus nancymae and Sapajus macrocephalus during three sexual phases (follicular, luteal, and gestational). Follicular quantification and immunolocalization of Caspase-3 protein, B-cell lymphoma 2 (BCL-2), proliferating cell nuclear antigen (PCNA), and anti-Mullerian hormone (AMH) were performed. A significant difference in the quantification between preantral and antral follicles, with a progressive decrease in the antrals, was identified. Protein and hormonal markers varied significantly between follicle cell types (A. nancymae p = 0.001; S. macrocephalus, p = 0.002). Immunostaining in the preantral and antral follicles was present in all sexual phases; for Caspase-3, in granulosa cells, oocytes, and stroma; for BCL-2, in granulosa cells, oocytes, and theca; and for PCNA and AMH, in oocytes and granulosa cells. The immunostaining for Caspase-3 was more expressive in the preantral follicles (follicular phase, p < 0.05), while that for BCL-2 and PCNA was more expressive in the antral follicles of the follicular phase. The AMH was more expressive in the primary and antral follicles of nonpregnant females, in both the follicular and luteal phases. Our results contribute to understanding the ovarian follicular selection, recruitment, and degeneration of these species.

Use of wildlife as an alternative protein source: Collared peccary meat.

Knowledge on the nutritional and sensory characteristics of wild meat provides a better basis for its use as food, ultimately expanding studies in gastronomy areas and stimulating the development of new products. This review aims to present information on the use of wild meat, with a focus on collared peccaries (Pecari tajacu). The biological characteristics of collared peccaries are discussed, with a focus on the main aspects of their meat. Collared peccary meat has excellent nutritional value due to its high protein levels (18.25%), unsaturated fatty acids (51.6-57.8%), and tenderness, similar to other domestic animals, thereby stimulating the interest of a new product market. Despite the demand for this product, collared peccary meat is scarce and not readily available for commercialization. Further, public policies are needed to encourage the management of this species to add value to the development of a production chain.

Early puberty in short-haired Guinea pigs kept in laboratory animal facilities.

Lab animals, such as Guinea pigs (Cavia porcellus), are crucial for scientific development, as they play an important role in the development and quality control chain of vaccines and drugs distributed by the Brazilian public health system. Investigating their biological and physiological parameters is fundamental to raise and keep these animals, so the handling of the facilities that hold them can be updated whenever new information comes up, with the well-being of the animals and alignment with the 3 Rs in mind. In the search for understanding reproductive aspects of Guinea pigs, the present study had the main goal of studying puberty by means of estrous cycle analysis in short-haired Guinea pigs. Guinea pigs have a vaginal occlusive membrane that covers the vaginal orifice. Its rupture takes place gradually and naturally, moments before labor and during estrus. The present study followed 42 females as for the presentation of the vaginal occlusive membrane. Once the membranes ruptured spontaneously, a swab was collected to study vaginal cytology. Membrane rupture was observed in 39 females; six females showed membrane rupture with less than 21 days of age (17 to 21 days). Twenty-three females were characterized as being in estrus due to cytology showing a prevalence of anucleated superficial cells. One of these females was younger than 21 days old. The opening of the vaginal occlusive membrane took place most frequently in intervals between 17 and 18 days, and the membrane remained open between one and three consecutive days. It was possible to follow three cycles of membrane opening on six females. The present study showed the need to adapt handling guidelines for C. porcellus kept in research animal facilities. The early age of puberty imposes the need of separate the female daughters from their fathers at 16 days old.

Sexual dimorphism in the turtle Kinosternon scorpioides (Testudines: Kinosternidae) from Marajó Island, Brazilian Amazon / Dimorfismo sexual de la tortuga Kinosternon scorpioides (Testudines: Kinosternidae) en la isla de Marajó, Amazonia brasileña

Abstract Introduction: Morphometrics analysis is an efficient and low-cost technique used in studies of sexual dimorphism in turtles. Kinosternon scorpioides scorpioides, scorpion mud turtle, has a wide phenotypic variation, depending on the area of its occurrence. Objective: The objective of this work was to identify the anatomical sexual difference of K. s. scorpioides, adults and hatchlings, through morphometric analysis; and relate the weights of adult animals to environmental factors (temperature and rainfall) in Marajó Island, Brazil. Methods: The sample collection was carried out from March to September 2018, covering both the rainy season (January to July) and the dry season (August to December). For the biometric analysis, 95 adults and 21 hatchlings were used, in which the length and width of the carapace and plastron, height of the shell, and weight were measured (adults only). For the geometric morphometry analysis, 21 adults and 13 hatchlings were used, in which 27 coordinates of anatomical landmarks were inserted in each image of the carapace and 11 in the plastron. Hatchlings were sexed by histology which was enabled by the identification of the ovaries and testicles. Results: The results showed the existence of dimorphism in adults. The plastron and height were higher in females, which had a more rounded carapace than males. This characteristic may be related to the species' sexual strategy, where males impose copulation. Histologically, it was possible to identify the ovaries and testicles in the hatchlings, but there was no anatomical sexual difference, despite the tendency to differentiate in the analysis of carapace PCA. Conclusions: Sexual dimorphism in K. s. scorpioides may play an important role on its reproductive behavior, which is synchronized with environmental events. This fact suggests that the reproductive strategies of this species would be severely affected by changes in the ecosystem.

Resumen Introducción: La morfometría es una técnica eficiente y de bajo costo, utilizada en estudios de dimorfismo sexual en tortugas, mediante el marcado de puntos anatómicos. Este grupo incluye Kinosternon scorpioides scorpioides, con una amplia variedad fenotípica, dependiendo del área en la que se encuentra presente. Objetivo: Este estudio tuvo como objetivo identificar la diferencia sexual entre K. s. scorpioides, adultos y juveniles, a través del análisis morfométrico; y verificar la relación entre las medidas de peso para adultos y los factores ambientales (temperatura y precipitación) en la isla de Marajó, Brasil. Métodos: La recolecta se realizó de marzo a septiembre de 2018, cubriendo tanto la temporada de lluvias (enero a julio) como la estación seca (agosto a diciembre). Para el análisis biométrico, se utilizaron 95 animales adultos y 21 crías, en los que se midió la longitud y el ancho del caparazón y el plastrón, la altura del casco y el peso en adultos. Para la morfometría geométrica, se utilizaron 21 adultos y 13 juveniles, en los que se identificaron 27 coordenadas de puntos de referencia anatómicos en el caparazón y 11 en el plastrón. Los individuos recién eclosionados fueron sexados por histología de rutina, que permite identificar ovarios y testículos. Resultados: Los datos analizados mostraron que existe dimorfismo sexual en adultos. El plastrón y la altura fueron mayores en las hembras, que tenían un caparazón más redondeado que los machos. Esta característica puede estar relacionada con la estrategia sexual de la especie, donde los machos imponen la cópula. En los juveniles recién eclosionados fue posible identificar histológicamente los ovarios y los testículos, pero no hubo diferencias sexuales en relación con la morfometría corporal, a pesar de la tendencia a diferenciarse en el análisis de PCA de caparazón. Conclusiones: El dimorfismo sexual en K. s. scorpioides puede desempeñar un papel importante en su comportamiento reproductivo, que ocurre en sincronía con los eventos ambientales. Este hecho sugiere que las estrategias reproductivas de esta especie se verían gravemente afectadas por los cambios en el ecosistema.

Reproductive biology of owl (Aotus spp.) and capuchin (Sapajus spp.) monkeys.

Studies related to morpho-physiology improve the understanding of biological mechanisms in non-human primates (NHP), contributing to research in several areas, especially those of assisted reproduction and biomedicine. The genera Aotus and Sapajus are important considerations in conducting studies related to reproductive biology and biomedical research, where animals of these genera can serve as models for species with vulnerable conservation status. The reproductive functions of animals of these genera can be used in conducting studies with other NHP species, for which biological material is difficult to access. The aim in conducting the current review was to compile the scientific literature on the biology and reproductive aspects of these two genera, to provide a for a greater understanding of these topics. In addition, a systematic review of literature allows for efficient access relevant information from an extended timeframe collated in a single document. There was, therefore, a systematic search for keywords based on reproductive studies with animals in these genera conducted, and 58 articles, 12 books, and three academic theses were subsequently selected. Even though there have been advances in knowledge of the reproductive biology of animals from these genera that have occurred in recent decades, especially research conducted when these NHP were in captivity, a wide variety of areas remain to be studied so that the understanding of reproduction in these NHP can be enhanced and the knowledge gained from these studies utilized for greater understanding of reproduction of NHP in general.

Reproductive monitoring of collared peccary females (Pecari tajacu) by analysis of fecal progesterone metabolites.

The measurement of fecal progesterone metabolites (fPM) by enzyme immunoassay analysis is a non-invasive technique that permits gathering reproductive information from wildlife without the stress associated with restraint. In the collared peccary (Pecari tajacu), a high correlation between serum progesterone and fPM levels (r2 = 0.783) suggests that fPM can be used to monitor their reproductive function. We monitored fPM during the estrous cycle of 15 collared peccary females. Estrous cycles averaged 27.9 ±â€¯4.5 days (n = 28), ranging from 21 to 36 days. The luteal phase was 22.2 ±â€¯4.8 days and the inter-luteal phase was 4.3 ±â€¯1.4 days. Mean concentration of fPM across pregnancy were not different from those found during the luteal phase (1230 ±â€¯718 and 1265 ±â€¯584 ng/100 mg dried feces, respectively), however, significant differences were found when luteal phase concentrations were compared only against fPM concentrations during late pregnancy. In addition, late pregnancy fPM concentrations (1893 ±â€¯551 ng/100 mg) were also significantly higher than those in the early (639 ±â€¯339 ng/100 mg) and mid (1134 ±â€¯449 ng/100 mg) pregnancy. For females during the early post-partum period, fPM concentrations were significantly increased (243 ±â€¯118 ng/100 mg) than those of non-cycling females (103 ±â€¯89 ng/100 mg). The analysis of fPM is a simple, non-invasive methodology to detect the ovarian activity in the collared peccary; moreover, it provides a husbandry tool, which may be used to help understand how social structure may impact reproduction.

Postnatal testis development in the collared peccary (Tayassu tajacu), with emphasis on spermatogonial stem cells markers and niche.

Collared peccaries (Tayassu tajacu) present a unique testis cytoarchitecture, where Leydig cells (LC) are mainly located in cords around the seminiferous tubules (ST) lobes. This peculiar arrangement is very useful to better investigate and understand the role of LC in spermatogonial stem cells (SSCs) biology and niche. Recent studies from our laboratory using adult peccaries have shown that the undifferentiated type A spermatogonia (Aund or SSCs) are preferentially located in ST regions adjacent to the intertubular compartment without LC. Following these studies, our aims were to investigate the collared peccary postnatal testis development, from birth to adulthood, with emphasis on the establishment of LC cytoarchitecture and the SSCs niche. Our findings demonstrated that the unique LC cytoarchitecture is already present in the neonate peccary's testis, indicating that this arrangement is established during fetal development. Based on the most advanced germ cell type present at each time period evaluated, puberty (the first sperm release in the ST lumen) in this species was reached at around one year of age, being preceded by high levels of estradiol and testosterone and the end of Sertoli cell proliferation. Almost all gonocytes and SSCs expressed Nanos1, Nanos2 and GFRA1. The analysis of SSCs preferential location indicated that the establishment of SSCs niche is coincident with the occurrence of puberty. Taken together, our findings reinforced and extended the importance of the collared peccary as an animal model to investigate testis function in mammals, particularly the aspects related to testis organogenesis and the SSCs biology and niche.

Fetal development of the Poeppig's woolly monkey (Lagothrix poeppigii).

The intrauterine development is an evolutionary strategy that prepares the neonate for extra-uterine life, thus providing important information on the life history of species. In this study, we described the external and internal morphology of 25 fetuses of Poeppig's woolly monkeys (Lagothrix poeppigii) by taking advantage of a 10-year participatory collection of biological samples originated from animals hunted for subsistence purposes in the Peruvian Amazon. Logistic regressions estimated the probability of occurrence of each external morphological characteristic in relation to the crown-rump length (CRL). The presence of nails, closed eyelids, differentiated genitalia and formed limbs with separation of the digits were observed in all analyzed fetuses (≥4.2 cm CRL). The other characteristics appeared in the following order: skin with epidermal pigmentation, oral and nasal mucosal pigmentation, tactile pelage and covering pelage. Although advanced fetuses (>15.8 cm CRL) showed most fetal external characteristics, they were not fully developed and no specimen showed tooth eruption or opened eyelids. The growth formula used to determine fetal age was ∛W = 0.042 (t - 45), with a high linear relationship between CRL and gestational age. All associations between the external biometry, absolute volume of internal organs and the CRL had a high coefficient of determination. Advanced fetuses and adults showed similar relative volume of thoracic and abdominal organs, except for thymus and the liver with a higher and lower relative volume, respectively. The relative volume of the tubular gastrointestinal tract and the thymus had a constant increase along fetal development, and the liver showed a significant decrease. This study describes important morphological events for understanding the gestational development in the Lagothrix genus. In addition, these results may be useful to improve imaging techniques, contributing to the in situ and ex situ reproductive management of this highly hunted species in the Amazon.

Embryonic and fetal morphology in the lowland paca (Cuniculus paca): A precocial hystricomorph rodent.

In mammals, the embryonic and fetal development of a species has evolved to maximize neonatal survival. In this study, we use a sample of 132 embryos/fetuses of wild lowland paca (Cuniculus paca), obtained over a period of 15 years through collaborative methods with local hunters in the Amazon to describe the intrauterine development of external and internal morphology of this Neotropical rodent. We also compare the newborn survival strategy in this species with other rodents. The crown-rump length (CRL) ranged between 0.6 and 24.6 cm. External features appeared in the following chronological order: limbs, eyelid buds, fusioned eyelids, genitalia, outer ear, tactile pelage, claws, skin, skin spots, covering pelage, teeth and open eyelids. Fetuses with CRL >19.5 cm presented all external features fully developed. The growth formula of fetal age was calculated as ∛W = 0.082 (t - 37.25), and age was accurately associated with CRL. We described the relationship between CRL and external and internal biometry. The liver declined in proportion within the internal cavity, while the relative volume of tubular gastrointestinal organs increased significantly along the embryo/fetal development. All organs, except the heart and the thymus, had similar relative volumes in advanced fetuses and adults. Our comparison of the intrauterine development in several rodent species indicates that the paca's reproductive strategy is comparable to species that are subject to low natural predation. Given that C. paca is perhaps the most hunted animal in Latin America, sustainable hunting throughout its range must take into account its relative reproductive performance.

Profile of plasmatic progesterone on pregnancy, and the postpartum estrus of Dasyprocta prymnolopha (Rodentia: Dasyproctidae) / Perfil de la progesterona plasmática durante durante la preñez y celo posparto del Dasyprocta prymnolopha (Rodentia: Dasyproctidae)

Abstract:The agouti (Dasyprocta sp.) is a hystricomorph rodent found in some regions of the Americas. It is an important cynegetic species, which indicates that the overhunting is a threat to their conservation. Very little is known about this wildlife in relation to what already has been studied in domestic animals. Thus, the knowledge on reproduction of wildlife becomes necessary and essential for the management and conservation of these natural resources. Specifically, studies regarding hormonal monitoring are important as a basic tool for research in modern reproductive biotechnology, and currently, there is no information on the progesterone changes during pregnancy of Dasyprocta sp., compared to other hystricomorphs. The aim of this study was to describe the profile of plasmatic progesterone during pregnancy, and report the restart of ovarian cycle in agouti after parturition. For this purpose, 18 black-rumped agoutis (Dasyprocta prymnolopha) born in captivity were used, and one or more consecutive pregnancies were observed. Copulation was confirmed by the presence of spermatozoa observed in the colpocytological examination. Blood samples were collected two times per week, and concentrations of progesterone, assessed in ten agoutis, were determined by radioimmunoassay. The onset of ovarian activity in six females was observed daily by colpocytological examination, starting on the seventh day postpartum. The gestational period observed in this study was 104.04 days (SD = 1.31) (101-106 days) (Number of cases, N = 26), and the interval between births was 126.03 days (SD = 18.40) (109-184 days). The plasmatic profile of progesterone during pregnancy showed a progressive increase from the 1st to the 5th week. The higher progesterone levels over this period (6.88 ng / mL, SD = 3.01) were detected in the 5th week. This value was similar (One-Way ANOVA, p > 0.05) to that observed in the 4th and 6th weeks, but was statistically different (One-Way ANOVA, p < 0.05) when compared to the other weeks. After the 6th week there was a progressive decrease in plasmatic progesterone levels. The animals showed a postpartum estrus of 12.04 days (SD= 4.29) (7-24 days) (N= 23). It was observed that 80.95 % (N= 19) of copulations during this period were fertile. This work contributed to understanding the dynamic changes in the progesterone levels during the pregnancy in agouti. Nevertheless, more studies are needed for a better appreciation of other endocrine and biological changes, in the mother and feto-placental unit of the agouti. Rev. Biol. Trop. 64 (4): 1519-1526. Epub 2016 December 01.

Resumen:El agouti (Dasyprocta sp.) es un roedor histricomorfo encontrado en algunas regiones de América. Es una especie cinegética importante, lo que indica que la caza excesiva es una amenaza para su conservación. Muy poco es lo que se conoce acerca de la biología de los animales silvestres, en relación al conocimiento acumulado sobre los domésticos. Por lo tanto, el estudio sobre la reproducción de los animales silvestres se hace necesario e imprescindible para el manejo y conservación de la especie como recurso natural. En concreto, los estudios relativos a la monitorización hormonal son una herramienta básica para la investigación en biotecnología reproductiva moderna. No hay información sobre los cambios de progesterona durante la preñez de Dasyprocta sp. en comparación con otros histricomorfos. El objetivo de este estudio fue describir el perfil de la progesterona plasmática durante la preñez, y reportar el reinicio del ciclo ovárico después del parto. Para ello, se utilizaron 18 agoutis de rabo negro (Dasyprocta prymnolopha) nacidos en cautiverio. Fueron estudiadas una o más preñeces consecutivas. La copulación fue confirmada por la presencia de espermatozoides observados en examen colpocitológico. Las muestras de sangre se recogieron dos veces por semana, y las concentraciones de progesterona se determinaron en 10 individuos mediante radioinmunoensayo. Se observó el inicio de la actividad ovárica diaria por examen colpocitológico, comenzando en el día séptimo posparto. El período gestacional observado en este estudio fue de 104.04 días (SD= 1.31, rango entre 101-106 días, N= 26), y el intervalo entre los nacimientos fue 126.03 días (SD= 18.40, rango entre 109-184 días). El perfil plasmático de la progesterona durante el embarazo mostró un aumento progresivo de la 1ª a la 5ª semana. Los mayores niveles de progesterona durante este período (6.88 ng / mL, SD= 3.01) se detectaron en la quinta semana. Este valor fue similar (ANOVA de um factor, p > 0.05) al observado en las semanas cuatro y seis, pero fue estadísticamente diferente (ANOVA de um factor, p<0.05) en comparación con las otras semanas. Después de la sexta semana hubo una disminución progresiva en los niveles plasmáticos de progesterona. Los animales mostraron un estro posparto de 12.04 días (SD= 4.29, rango entre 7-24 días, N= 23). Se observó que el 80.95 % (N= 19) de cópulas durante este período fueron fértiles. Este trabajo contribuyó a la comprensión de los cambios dinámicos en los niveles de progesterona durante la preñez del agouti. Sin embargo, se necesitan más estudios para una mejor comprensión de otros cambios biológicos y endocrinos, y en la unidad materna y feto-placentaria del agouti.

Serological evaluation of ovarian steroids of red-rumped agouti (Dasyprocta leporina) during the estrous cycle phases.

The objective of this study was to evaluate serum progesterone (P4) and 17ß estradiol (E2) concentrations throughout the estrous cycle in the red-rumped agouti (Dasyprocta leporina). A total of eight multiparous, captive-bred females were bled throughout their estrous cycle via saphenous venipuncture, with E2 and P4 concentrations being measured via ELISA of the serum collected. Serum E2 ranged from 1212 to 3500pg/ml with a peak value coinciding with observed estrus. However, two additional peak values for E2 were also recorded, one each in metestrus and proestrus respectively. P4 concentrations reached a maximum of 4.23ng/ml, and increases in P4 immediately followed the second E2 peak in metestrus. The highest concentrations of P4 were recorded in mid diestrus; between days 23-25 of the 31-day cycle. This phase was the longest in the agouti, consisting of approximately 19days and accounted for 61% of the cycle. This study increased the basal scientific reproductive knowledge of this potentially valuable neo-tropical species.

Sexual behavior of Pecari tajacu (Cetartiodactyla: Tayassuidae) during periovulatory and early gestation periods.

The goal of this study was to describe the sexual behavior in female and male collared peccary. Twenty females and twelve males were monitored in familiar and non-familiar units for two 60 days periods. During both phases, we recorded 2747 sexual interactions initiated by 20 different females toward males and 4461 sexual interactions initiated by 12 males toward females. The frequency of sexual interactions initiated per female significantly increased from proestrus to estrus, and they were significantly more frequently courted. Females initiated olfactory inspections 15.42 times more and were mounted 22.6 times more during estrus than during proestrus. Nulliparous and primiparous females copulated only when exposed to non-parental males. After estrus, the frequency of sexual interactions received by females sharply decreased. One mating event was recorded during the first gestation week and 31 mountings were observed after the second week. In conclusion, the behavioral monitoring is a useful procedure for the recognition of estrus. Our results suggest that ovulation may be associated with the end of the estrus, which will support future work in assisted reproduction in this species. To promote good handling practices, females of reproductive age should be removed from their family unit of origin.

Profile of plasmatic progesterone on pregnancy, and the postpartum estrus of Dasyprocta prymnolopha (Rodentia: Dasyproctidae).

The agouti (Dasyprocta sp.) is a hystricomorph rodent found in some regions of the Americas. It is an important cynegetic species, which indicates that the overhunting is a threat to their conservation. Very little is known about this wildlife in relation to what already has been studied in domestic animals. Thus, the knowledge on reproduction of wildlife becomes necessary and essential for the management and conservation of these natural resources. Specifically, studies regarding hormonal monitoring are important as a basic tool for research in modern reproductive biotechnology, and currently, there is no information on the progesterone changes during pregnancy of Dasyprocta sp., compared to other hystricomorphs. The aim of this study was to describe the profile of plasmatic progesterone during pregnancy, and report the restart of ovarian cycle in agouti after parturition. For this purpose, 18 black-rumped agoutis (Dasyprocta prymnolopha) born in captivity were used, and one or more consecutive pregnancies were observed. Copulation was confirmed by the presence of spermatozoa observed in the colpocytological examination. Blood samples were collected two times per week, and concentrations of progesterone, assessed in ten agoutis, were determined by radioimmunoassay. The onset of ovarian activity in six females was observed daily by colpocytological examination, starting on the seventh day postpartum. The gestational period observed in this study was 104.04 days (SD = 1.31) (101-106 days) (Number of cases, N = 26), and the interval between births was 126.03 days (SD = 18.40) (109-184 days). The plasmatic profile of progesterone during pregnancy showed a progressive increase from the 1st to the 5th week. The higher progesterone levels over this period (6.88 ng / mL, SD = 3.01) were detected in the 5th week. This value was similar (One-Way ANOVA, p > 0.05) to that observed in the 4th and 6th weeks, but was statistically different (One-Way ANOVA, p < 0.05) when compared to the other weeks. After the 6th week there was a progressive decrease in plasmatic progesterone levels. The animals showed a postpartum estrus of 12.04 days (SD= 4.29) (7-24 days) (N= 23). It was observed that 80.95 % (N= 19) of copulations during this period were fertile. This work contributed to understanding the dynamic changes in the progesterone levels during the pregnancy in agouti. Nevertheless, more studies are needed for a better appreciation of other endocrine and biological changes, in the mother and feto-placental unit of the agouti.

Epididymal morphophysiology of adult collared peccaries (Pecari tajacu) / Morfofisiologia epididimária de catitus (Pecari tajacu) adultos

A functional understanding of the epididymis makes it possible to increase a species' fertility, since it facilitates the preservation of gametes for use in assisted reproduction techniques. This study analyzed the histological features of the different regions of the epididymis of the Pecari tajacu and the associated pathologies found in the spermatozoa present in this organ. After an orchiectomy surgery to remove the testicles and epididymis obtained from eight adult animals bred in captivity, fixation and histological processing of samples were carried out. The presence of pseudostratified columnar epithelium with stereocilia was observed in all regions of the epididymis (caput, corpus and caudal). Similarities were noted between the average height of the corpus and caudal epithelium, which differed from that found at the caput (1- proximal: region associated with the efferent ducts; 2- distal: region associated with corpus epididymis). It was also observed that the proximal caput presents a smaller average of the tubules diameter compared to other portions of the epididymis. Regarding pathologies of the spermatozoa, thirteen different types of morphological modifications were observed. Underdeveloped spermatozoa and detached heads were most frequently encountered along the epididymis of the collared peccaries. This study is a pioneer in the area and its data will serve as a basis for comparative research on the reproductive biology of artiodactyls, thus contributing to their conservation and reproduction.(AU)

O entendimento funcional do epidídimo possibilita o aumento da fertilidade de uma espécie, pois torna possível a preservação de gametas para o uso em técnicas de reprodução assistida. Este trabalho estudou os aspectos histológicos das diferentes regiões do epidídimo de Pecari tajacu, bem como as patologias associadas encontradas nos espermatozoides presentes nesse órgão. Após procedimento cirúrgico de orquiectomia, para remoção dos testículos e epidídimos, obtidos de oito animais adultos nascidos em cativeiro, realizou-se a fixação e o processamento histológico das amostras. Constatou-se em todas as regiões do epidídimo (cabeça, corpo e cauda) a presença de epitélio pseudoestratificado colunar com estereocílios. Foram observadas semelhanças entre a altura média do epitélio do corpo e da cauda, a qual era diferente da encontrada na cabeça (1- proximal: região associada aos ductos eferentes; 2- distal: região associada ao corpo do epidídimo). Foi verificado que a cabeça proximal apresenta a média de diâmetro dos túbulos menor em relação às demais porções do epidídimo. Em relação às patologias dos espermatozoides, constataram-se treze tipos diferentes de alterações morfológicas. Os espermatozoides subdesenvolvidos e cabeças soltas foram as mais frequentes ao longo do epidídimo de catitus. Os dados deste trabalho são pioneiros, e servirão como base para pesquisas comparativas sobre a biologia reprodutiva dos artiodátilos, contribuindo para a sua conservação e produção.(AU)

Derivation of sperm from xenografted testis cells and tissues of the peccary (Tayassu tajacu).

Because the collared peccary (Tayassu tajacu) has a peculiar Leydig cell cytoarchitecture, this species represents a unique mammalian model for investigating testis function. Taking advantage of the well-established and very useful testis xenograft technique, in the present study, testis tissue and testis cell suspensions from immature collared peccaries (n=4; 3 months old) were xenografted in SCID mice (n=48) and evaluated at 2, 4, 6, and 8 months after grafting. Complete spermatogenesis was observed at 6 and 8 months after testis tissue xenografting. However, probably due to de novo testis morphogenesis and low androgen secretion, functionally evaluated by the seminal vesicle weight, a delay in spermatogenesis progression was observed in the testis cell suspension xenografts, with the production of fertile sperm only at 8 months after grafting. Importantly, demonstrating that the peculiar testicular cytoarchitecture of the collared peccary is intrinsically programmed, the unique Leydig cell arrangement observed in this species was re-established after de novo testis morphogenesis. The sperm collected from the xenografts resulted in diploid embryos that expressed the paternally imprinted gene NNAT after ICSI. The present study is the first to demonstrate complete spermatogenesis with the production of fertile sperm from testis cell suspension xenografts in a wild mammalian species. Therefore, due to its unique testicular cytoarchitecture, xenograft techniques, particularly testis cell suspensions, may represent a new and very promising approach to evaluate testis morphogenesis and to investigate spermatogonial stem cell physiology and niche in the collared peccary.

Puberty in male collared peccary (Pecari tajacu) determined by quantitative analysis of spermatogenic cells / Puberdade em macho de cateto (Pecari tajacu) determinada pela análise quantitativa das células espermatogênicas

Biological studies are necessary for the management of wildlife in captivity, and knowledge of reproduction is one of the important features for increasing production. The objective of the research was to determine the age at which male collared peccaries reach puberty. Testicular samples of 15 animals, aged 7 to 16 months, distributed into five groups (G1, G2, G3, G4 and G5) were used. The testes showed considerably increased weight, length and width (p < 0.05) from G1 to G3, whereas, from this group onward, the development of this organ was slower. There was positive correlation (p < 0.001) between the following testicular parameters: weight and length (r = 0.97), weight and width (r = 0.88), length and width (r = 0.92). Regarding the diameter of seminiferous tubules, an increase was observed (p < 0.05) from G1 to G4. The total number of spermatogenic cells increased significantly (p < 0.05) until G3 and then it stabilized. There was also positive correlation between testis weight and tubular diameter (r = 0.99, p < 0.001), and testis weight and spermatogenic cells (r = 0.98, p < 0.001). The number of Sertoli cells decreased significantly (p < 0.05) from G1, when they were undifferentiated as support cells, to G5, when they occurred together with the complete line of spermatic cells. The results demonstrate that the reproductive development of peccaries can be classified into the following stages: impuberty (G1, 7-8 months); pre-pubertal (G2, 9-10 months); pubertal (G3, 11-12 months); post-pubertal 1 (G4, 13-14 months); and post-pubertal 2 (G5, 15-16 months). Based on the histological analyses, puberty in the male collared peccary was determined to occur between 11 and 12 months of age.

Estudos biológicos são necessários para o manejo da vida silvestre em cativeiro, e o conhecimento da reprodução é um dos aspectos importantes para o aumento da produção. Esta pesquisa teve como objetivo determinar a idade da puberdade do cateto macho. Foram utilizadas amostras testiculares de 15 animais, entre 7 a 16 meses, distribuídos em cinco grupos (G1, G2, G3, G4 e G5). Os testículos aumentaram no peso, comprimento e largura consideravelmente (p < 0,05) do G1 ao G3, enquanto que, a partir deste grupo, o desenvolvimento desse órgão foi mais lento. Houve correlação positiva (p < 0,001) entre os seguintes parâmetros testiculares: peso e comprimento (r = 0,97), peso e largura (r = 0,88), comprimento e largura (r = 0,92). Com relação ao diâmetro tubular, observou-se um aumento (p < 0,05) do G1 ao G4. A quantidade total de células espermatogênicas aumentou significativamente (p < 0,05) até o G3, e se estabilizou a partir deste grupo. Houve correlação positiva entre o peso testicular e o diâmetro tubular (r = 0,99, p < 0,001), bem como o peso testicular e as células espermatogênicas (r = 0,98, p < 0,001). A quantidade de células de Sertoli reduziu significativamente (p < 0,05) do G1, onde se encontravam indiferenciadas como células de suporte, até G5, onde foram observadas juntamente com todas as células da linhagem espermática. Estes resultados demonstraram que as fases do desenvolvimento reprodutivo de catetos podem ser classificadas em: impúbere (G1, 7-8 meses), pré-púbere (G2, 9-10 meses), púbere (G3, 11-12 meses), pós-púbere 1 (G4, 13-14 meses) e pós-púbere 2 (G5, 15-16 meses). Com base na análise histológica, a puberdade dos catetos machos ocorre entre 11 e 12 meses de idade.

Progesterone and estradiol-17ß as a potential method for pregnancy diagnosis in the collared peccary (Pecari tajacu).

In this study, the period of pregnancy of nine collared peccary females has been monitored through the analysis of serum progesterone and estradiol-17ß profiles. Serum concentrations of progesterone increased by Day 4 after conception, reaching concentrations of 33.4±5.6 ng/mL on Day 10. Between Days 10 and 130 progesterone values were maintained between 20 and 60 ng/mL. In the collared peccary, embryonic estradiol synthesis is first observed in the systemic circulation by Day 15 of pregnancy. Between Days 0 and 50 of pregnancy, average estradiol-17ß concentrations were between 0 and 30 pg/mL. From Day 75 of pregnancy onwards, estradiol concentrations were constantly increasing, reaching maximum concentrations (131.4±40.8 pg/mL) on the day of parturition. The combined study of serum progesterone and estradiol-17ß concentrations as a potential method for early pregnancy diagnosis presented the best overall accuracy (73%) when the threshold was established at 20 ng/mL serum progesterone and 20 pg/mL serum estradiol. Nevertheless, the accuracy for diagnosing pregnancy of females at mid and late pregnancy was 78% and 95%, respectively. The analysis of the sexual hormones during pregnancy could be a useful tool as a potential pregnancy diagnosis and an efficient predictor of the day of parturition in the captive collared peccary.

Ovarian folliculogenesis in collared peccary, Pecari tajacu (Artiodactyla: Tayassuidae).

The sustainability and production of collared peccary (Pecari tajacu) has been studied in the last few years; however, further information on its reproduction is necessary for breeding systems success. Understanding folliculogenesis aspects will contribute to effective reproductive biotechniques, which are useful in the preservation and production of wildlife. The aim of this study was-to evaluate the ovarian folliculogenesis in collared peccary. Ovaries from six adult females of collared peccary were obtained through ovariectomy and analyzed. These were fixed in aqueous Bouin's solution and sectioned into 7 microm slices, stained with hematoxilin-eosin and analyzed by light microscopy. The number of pre-antral and antral follicles per ovary was estimated using the Fractionator Method. The follicles, oocytes and oocyte nuclei were measured using an ocular micrometer. Results showed that the length, width, thickness, weight, and the gross anatomy of the right and left ovaries were not significantly different. However, the mean number of corpora lutea was different between the phases of the estrous cycle (p<0.05), with the highest mean in the luteal phase. Primordial follicles were found in the cortex; the oocytes were enveloped by a single layer of flattened follicular cells. In the primary follicles, proliferation of the follicular cells gave rise to cuboidal cells (granulosa cells). The secondary follicle was characterized by two or more concentric layers of cuboidal cells (granulosa), beginning of antrum formation, and the presence of pellucid zone and theca cells. Antral follicles were characterized by a central cavity (antrum), the presence of cumulus oophorus and theca layers (interna and externa). In the right ovary, the values of the primordial and primary follicles were similar, but significantly different from the secondary ones (p<0.05). In the left ovary, significant differences were observed between all follicles in the follicular phase (p<0.05); the mean number of primordial and primary follicles was similar in the luteal phase. The mean number of pre-antral follicles and antral follicles in the follicular phase was higher in the left ovary (p<0.05). The mean number of antral follicles in the luteal phase was similar in both ovaries. We also found significant differences in mean diameter of preantral follicles, oocyte, granulosa layer and oocyte nucleus during the estrous cycle. In the antral follicles a significant difference was observed only in follicular diameter (p<0.05). The predominance of active primordial and primary follicles was found in both phases; otherwise the secondary follicles and antral follicles showed a high degree of degeneration. The results obtained in the present work will strengthen the development of biotechnology programs to improve the productive potential and conservation of the collared peccary.

Ovarian folliculogenesis in collared peccary: Pecari tajacu (Artiodactyla: Tayassuidae)

The sustainability and production of collared peccary (Pecari tajacu) has been studied in the last few years; however, further information on its reproduction is necessary for breeding systems success. Understanding folliculogenesis aspects will contribute to effective reproductive biotechniques, which are useful in the preservation and production of wildlife. The aim of this study was to evaluate the ovarian folliculogenesis in collared peccary. Ovaries from six adult females of collared peccary were obtained through ovariectomy and analyzed. These were fixed in aqueous Bouin’s solution and sectioned into 7μm slices, stained with hematoxilin-eosin and analyzed by light microscopy. The number of pre-antral and antral follicles per ovary was estimated using the Fractionator Method. The follicles, oocytes and oocyte nuclei were measured using an ocular micrometer. Results showed that the length, width, thickness, weight, and the gross anatomy of the right and left ovaries were not significantly different. However, the mean number of corpora lutea was different between the phases of the estrous cycle (p<0.05), with the highest mean in the luteal phase. Primordial follicles were found in the cortex; the oocytes were enveloped by a single layer of flattened follicular cells. In the primary follicles, proliferation of the follicular cells gave rise to cuboidal cells (granulosa cells). The secondary follicle was characterized by two or more concentric layers of cuboidal cells (granulosa), beginning of antrum formation, and the presence of pellucid zone and theca cells. Antral follicles were characterized by a central cavity (antrum), the presence of cumulus oophorus and theca layers (interna and externa). In the right ovary, the values of the primordial and primary follicles were similar, but significantly different from the secondary ones (p<0.05). In the left ovary, significant differences were observed between all follicles in the follicular phase (p<0.05); the mean number of primordial and primary follicles was similar in the luteal phase. The mean number of pre-antral follicles and antral follicles in the follicular phase was higher in the left ovary (p<0.05). The mean number of antral follicles in the luteal phase was similar in both ovaries. We also found significant differences in mean diameter of preantral follicles, oocyte, granulosa layer and oocyte nucleus during the estrous cycle. In the antral follicles a significant difference was observed only in follicular diameter (p<0.05). The predominance of active primordial and primary follicles was found in both phases; otherwise the secondary follicles and antral follicles showed a high degree of degeneration. The results obtained in the present work will strengthen the development of biotechnology programs to improve the productive potential and conservation of the collared peccary.

La sustentabilidad y la producción de pecarí de collar (Pecari tajacu) han sido estudiados en los últimos años, sin embargo, más información sobre su reproducción es necesaria para el éxito de los sistemas de crianza . La comprensión de los aspectos relacionados con la foliculogénesis contribuirá con la aplicación de biotécnicas de reproducción, las cuales son útiles en la preservación y la producción de la vida silvestre. El objetivo de este estudio fue obtener datos sobre la población folicular del ovario de pecarí de collar. En relación con la población folicular en el ovario derecho, los valores de los folículos primordiales y primarios fueron similares, pero se observó que había una diferencia significativa (p<0.05) con el secundario. En el ovario izquierdo, la fase folicular presentó diferencias significativas (p<0.05) entre todos los folículos, y en la fase lútea el número medio de folículos primordiales y primarios fueron similares. Ahora bien, con respecto a la población de folículos antrales, en la fase folicular, se observaron diferencias significativas entre los ovarios (p<0.05), y de forma similar en la fase lútea. También se encontraron diferencias significativas en el diámetro medio de los folículos preantrales, oocitos, la capa granulosa y el núcleo del oocito durante las fases del ciclo estral. Asimismo, en los folículos antrales se observó diferencia estadísticamente significativa (p<0.05) en el diámetro folicular. En ambas fases del ciclo estral, se encontró el predominio de folículos primordiales y primarios en desarrollo, por otro lado, los folículos secundarios y los folículos antrales mostraron un alto grado de degeneración. Los resultados aquí presentes son necesarios para el desarrollo de los programas de mejoramiento y conservación.